Anyone know where to obtain beta amylase enzyme (without any other enzymes mixed in) for a reasonable price? I have an experiment I would like to do, but it wouldn't be complete if I couldn't get isolated beta amylase.
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Brew o
Beta amylase available anywhere?
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OP
OP
Bump.
I would be willing to work with beta mixed with alpha amylase, but no other enzymes.
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I would be willing to work with beta mixed with alpha amylase, but no other enzymes.
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DuncB
Well-Known Member
Well done for tracking that down I searched for a while but couldn't find the goods.
OP
OP
Thanks, but I've seen that listing. I'm not interested enough in the experiment to spend $300+ to run the experiment.
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Not sure what your budget is, but Fisher has 25g for $69.
https://www.fishersci.com/shop/products/beta-amylase-soybean-tci-america/A044825G
https://www.fishersci.com/shop/products/beta-amylase-soybean-tci-america/A044825G
OP
OP
Not sure what your budget is, but Fisher has 25g for $69.
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This is a lot more reasonable. Thank you.
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Clearly I saw the price.
I wonder if there's a difference between barley vs soybean product.
Given the huge price disparity I would make sure the TCI product is truly pure beta amylase...
Cheers!
I wonder if there's a difference between barley vs soybean product.
Given the huge price disparity I would make sure the TCI product is truly pure beta amylase...
Cheers!
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OP
OP
I might as well go ahead and outline the experiment I am looking at.
It is my belief, based on research, that it is really limit dextrinase, and not beta amylase, that is responsible for the higher fermentability of worts mashed at low initial temperatures compared to worts mashed at higher initial temperatures. So, I have an experiment in mind that could confirm or refute this hypothesis.
The basic outline is:
If the wort mashed with beta + alpha has a lower FG than the wort mashed only with alpha, then beta amylase action does in fact lead to higher fermentability than mashing with alpha alone. If the two worts finish at the same FG, then beta action does not actually do anything to increase the fermentability of worts. If the second case holds, then the higher fermentability of low mashed worts is due to something other than beta amylase.
Comments? @VikeMan
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It is my belief, based on research, that it is really limit dextrinase, and not beta amylase, that is responsible for the higher fermentability of worts mashed at low initial temperatures compared to worts mashed at higher initial temperatures. So, I have an experiment in mind that could confirm or refute this hypothesis.
The basic outline is:
- Denature all of the endogenous enzymes by doughing in to boiling water, and then conducting a cereal mash to fully gelatinize all of the starch in the grain.
- Cool the mash down to ~145°F (63°C) and maintain temp with a Sous Vide controller.
- Add either alpha amylase alone, or alpha + beta amylase to the mash, and let the mash go for ~2 hours to make absolutely sure that the enzymes have done all they possible can.
- Ferment both worts using a fast ferment test schedule (with the same yeast obviously.)
Comments? @VikeMan
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Red over White
Well-Known Member
My monies on limit dextrinase being imperative for highly fermentable wort. Budweiser doesn’t start their mash at 113°F for beta.
VikeMan
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Very interesting! My suggestions, some of which you are probably already planning, but omitted for brevity:
- Boil or otherwise sanitize/sterilize the worts before fermentation.
- Measure the OGs
- Use a yeast strain that's non-diastatic
- Consider repeating (if affordable) with one hour mashes and no cereal mash, to approximate normal/typical mash conditions. Could remove possible objections that could arise from only doing it with a cereal mash and longer mash. (I wouldn't personally predict the basic result to be different between the two trials, but I'd still probably do them.)
- If planning to pulverize the grains, do a dry run to make sure they don't tend to "brick" at the bottom of the mash vessel. I've seen that with some malts during pH experiments.
ETA: - Consider repeating with different total enzyme concentrations.
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OP
OP
I just purchased a vial of enzymes designed for the fermenter, to make a brut IPA/Lager. The LHBS had three different compounds, and I just bought the one he suggested, but I have no idea at the moment what % of which enzymes are in it or the recipes for the ones I didn't buy (one of which was mash-oriented). Are these compounds that difficult to find or is it that your parameters were very specific?
OP
OP
There are lots of useful enzymes and mix combos available. But for the purposes of this experiment, I need to specifically have beta amylase, and this is not commonly available.
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WestMichiganSteelheader
Well-Known Member
This is what I love about HB and this website. We science nerds love these ideas.I might as well go ahead and outline the experiment I am looking at.
It is my belief, based on research, that it is really limit dextrinase, and not beta amylase, that is responsible for the higher fermentability of worts mashed at low initial temperatures compared to worts mashed at higher initial temperatures. So, I have an experiment in mind that could confirm or refute this hypothesis.
The basic outline is:
If the wort mashed with beta + alpha has a lower FG than the wort mashed only with alpha, then beta amylase action does in fact lead to higher fermentability than mashing with alpha alone. If the two worts finish at the same FG, then beta action does not actually do anything to increase the fermentability of worts. If the second case holds, then the higher fermentability of low mashed worts is due to something other than beta amylase.
- Denature all of the endogenous enzymes by doughing in to boiling water, and then conducting a cereal mash to fully gelatinize all of the starch in the grain.
- Cool the mash down to ~145°F (63°C) and maintain temp with a Sous Vide controller.
- Add either alpha amylase alone, or alpha + beta amylase to the mash, and let the mash go for ~2 hours to make absolutely sure that the enzymes have done all they possible can.
- Ferment both worts using a fast ferment test schedule (with the same yeast obviously.)
Comments? @VikeMan
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What do you propose for a control? Are you going to split a batch with and without the enzyme from the same boil? Keep us posted with your progress.
If I was doing a super dry Brut I would use glucoamylase as that will rip virtually everything and end up at 1.000 fg...
Cheers!
Cheers!
OP
OP
Control would be a mash conducted normally at the ~145°F temp. If my hypothesis is correct, this wort would have higher fermentability, due to the presence of limit dextrinase, than either of the experimental cells.
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I think this is pretty much the only way to make a real Brut. Diastatic strains make their own glucoamylase, but I don't believe that they'll ferment down to 1.000 or below.
I guess you can't really do the perfect experiment since it's pretty much impossible to buy limit dextrinase. I suppose you could try making your own.
OP
OP
The extraction method is a little beyond my means, both equipment and skills. Will have to stick to the simple experiment.I guess you can't really do the perfect experiment since it's pretty much impossible to buy limit dextrinase. I suppose you could try making your own.
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Is your working hypothesis that limit dextrinase contributes to wort fermentability, especially in low-temperature mashes, or is it that beta amylase does not have a role? I don't think the first statement is at all controversial.
If it's the second, it would be interesting if you could provide more of your reasoning. In particular I'm wondering about high-temperature (75 C) mashing to produce wort with poor fermentability (for the low-alcohol folks), where the standard explanation is that you're eliminating beta amylase activity and as a result leaving behind longer 1,4-linked oligosaccharides.
It seems like your experiment will have to be careful to match the enzyme concentration and duration of a typical mash. Otherwise, you will only reach a weaker conclusion: "Given enough alpha amylase and enough time, you can achieve a similar fermentability without beta amylase," instead of "In a normal mash, beta activity does not contribute to starch hydrolysis."
If it's the second, it would be interesting if you could provide more of your reasoning. In particular I'm wondering about high-temperature (75 C) mashing to produce wort with poor fermentability (for the low-alcohol folks), where the standard explanation is that you're eliminating beta amylase activity and as a result leaving behind longer 1,4-linked oligosaccharides.
It seems like your experiment will have to be careful to match the enzyme concentration and duration of a typical mash. Otherwise, you will only reach a weaker conclusion: "Given enough alpha amylase and enough time, you can achieve a similar fermentability without beta amylase," instead of "In a normal mash, beta activity does not contribute to starch hydrolysis."
OP
OP
The hypothesis is that the higher fermentability of worts mashed initially at lower temperatures is due to the action of limit dextrinase, and that beta amylase does nothing to contribute to the higher fermentability (but does help alpha amylase with hydrolysis of starch.) Limit dextrinase is the only enzyme in barley malt that can hydrolyze the alphat(1-6) bonds that create branch points in amylopectin. By hydrolyzing some of the branch bonds, limit dextrinase reduces the amount of limit dextrin that will be left when alpha and beta amylase have hydrolyzed everything that they can. Less dextrin in the final wort leads to a more fermentable wort, since dextrins are not fermentable.
High initial mash temperatures can rapidly denature limit dextrinase, just like beta amylase. So, if you denature the limit dextrinase early, you will have more dextrin in the final wort, and it will be less fermentable because of that.
I never postulated that beta amylase does not contribute to starch hydrolysis. It does, and it makes the overall process of saccharification go faster. But since gelatinization is the rate limiting step in starch conversion, the speed of hydrolysis is a minor factor in time to "complete" a mash. What the experiment is designed to elucidate, is whether or not having beta working with alpha results in more fermentable wort - so, just to support your so called "weaker conclusion", and that beta is not responsible for the higher fermentability of worts initially mashed at low termperatures. To do this, we need to denature the limit dextrinase very early on in the mash, unfortunately, this will also denature the beta and alpha amylase, thus the need to add exogenous amylase enzymes for the experiment.
There is a great deal of misunderstanding of exactly how alpha amylase works. Statements I have seen many times are: "alpha amylase only produces unfermentable sugars", and the equivalent "alpha amylase does not produce fermentable sugars." Both of these are false. Alpha amylase can produce sugars of any size including mono- and di-saccharides, which are fermentable. What is true, is that early on in the hydrolysis process alpha produces mostly large, non-fermentable sugars. But the longer alpha acts the more fermentable sugars are produced, as the hydrolysis fragments get smaller and smaller with continued alpha action. This behavior is due to alpha amylase being able to hydrolyze alpha(1-4) bonds at random locations along starch chains.
Beta amylase is easier to understand, as it only creates maltose (a fermentable di-saccharide.) If you misunderstand alpha amylase as discussed in the previous paragraph, then it only seems logical that you need beta amylase to turn the unfermentable sugars created by alpha amylase into fermentable sugars. But, once you know how alpha actually works, then it's not so difficult to understand that you only need alpha to create fermentable sugars, but that having beta available speeds things up a bit.
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For a hot mash in the zone for alpha only, where the beta (and I suppose the limit dextrinase also) have been denatured, I have figured one can obtain a reasonably fermentable wort anyway simply by mashing for a long time, such as with an overnight mash of 6-10 hours or whatever. I've yet to experiment with this hypothesis.
What many do not realize is that there are no magical temperatures at which the enzymes decide to start working and then stop working. The enzymes are more like little machines that run slower or faster based on temperature. Even at room temperature these little machines are moving, just very slowly. When temperatures get hot enough, the machines are running too fast and begin to break. Some of their brethren continue on for a while, as more and more of their other brethren fail. But it's not like they all get wiped out instantly unless the temperature rise is extremely severe.
(And yes I realize that gelatinization also plays an important role.)
(Moisture also plays a very important role. Dry enzymes can withstand higher temperatures. Otherwise kilned malts would not be very diastatic! I do hypothesize also that the enzymes in barely-crushed malt can allow for slow release of enzymes -- any portions of kernels that are not seeing much moisture until later on in the mash can handle higher temps for longer.)
What many do not realize is that there are no magical temperatures at which the enzymes decide to start working and then stop working. The enzymes are more like little machines that run slower or faster based on temperature. Even at room temperature these little machines are moving, just very slowly. When temperatures get hot enough, the machines are running too fast and begin to break. Some of their brethren continue on for a while, as more and more of their other brethren fail. But it's not like they all get wiped out instantly unless the temperature rise is extremely severe.
(And yes I realize that gelatinization also plays an important role.)
(Moisture also plays a very important role. Dry enzymes can withstand higher temperatures. Otherwise kilned malts would not be very diastatic! I do hypothesize also that the enzymes in barely-crushed malt can allow for slow release of enzymes -- any portions of kernels that are not seeing much moisture until later on in the mash can handle higher temps for longer.)
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DuncB
Well-Known Member
@doug293cz
Will there be spontaneous breakdown of the sugars in the warm Liquor?
The enzyme just catalyses the reaction to get the conversion in a suitable time I suppose.
Should be a slow constant for the experiment?
EDIT I see @dmtaylor was on this ahead of me.
Will there be spontaneous breakdown of the sugars in the warm Liquor?
The enzyme just catalyses the reaction to get the conversion in a suitable time I suppose.
Should be a slow constant for the experiment?
EDIT I see @dmtaylor was on this ahead of me.
The estimates I’ve found are that once the amylolytic enzymes have broken all the 1,4 bonds they can, the little 1,6-branched pieces that are left comprise about 10-15% of the total starch.
If alpha amylase alone is sufficient to get you all the way to limit dextrins, you’d expect an alpha-only mash to be ~85% fermentable sugars. High-temperature mashing gives you wort that’s <50% fermentable.
If alpha amylase alone is sufficient to get you all the way to limit dextrins, you’d expect an alpha-only mash to be ~85% fermentable sugars. High-temperature mashing gives you wort that’s <50% fermentable.
Indeed, I would expect this to come pretty close to reality. Not sure what a diastaticus yeast like 3711 or Belle might be able to do here though.
I am not certain but have the feeling that this the very low end of the fermentability range for any hot mash of any starch. Even bread & water with no added sugars will somehow ferment to about 50% AA. Ever heard of kvass? It is one of the strangest "beers" I know of, fermented bread-water. I made it once. Some of the starch and dextrins seem to be able to be broken down enough via whatever process (I'm not certain how) to allow partial fermentability. There's probably just too many variables for me to be able to understand how this works -- a little sugar in the bread, a little enzymatic action within the yeast cells, the temperatures involved breaking down some dextrins into actual fermentable sugars... boggles my mind somewhat that yeast is still able to eat a large percentage of what should be not far from 100% complex starch.
Ferment everything, all the way down. The glucoamylase produced by diastatic yeasts will break all starch down into simple sugars. (Non-starch carbohydrates, like beta glucans, are not broken down.)
The last beer I made mashed at 167 F went from 1.025 to 1.016 for 36% apparent attenuation.
OP
OP
If this were the case, then boiling wort should work even faster to hydrolyze starch to sugar, since the temp is higher, and reactions go faster the higher the temperature (all else being equal.)
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OP
OP
What do you expect if you mash at mashout temperatures? The alpha will be very short lived at that temp, so it's cumulative activity will be quite low. And due to the random cleave point action of alpha amylase, little in the way of fermentable sugar will be produced - the alpha just doesn't last long enough to get anywhere near the max fermentability point.
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OP
OP
But then, wort extract is only about 90% carbohydrates, so if 85% of that 90% is fermentable, then the fermentability is about 76-77%. Apparent attenuation will be different than that since the presence of alcohol reduces the SG.
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beren
Well-Known Member
Isn’t it beano?Anyone know where to obtain beta amylase enzyme (without any other enzymes mixed in) for a reasonable price? I have an experiment I would like to do, but it wouldn't be complete if I couldn't get isolated beta amylase.
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VikeMan
It ain't all burritos and strippers, my friend.
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No. Beano is Alpha Galactosidase.
huckdavidson
Well-Known Member
India might be a good source to investigate for beta and/or alpha amylase.
125 g for ~16 usd + shipping.
https://onlineingredient.com/produc...-amylase-enzyme-powder?variant=41597769285677
Or a more expensive alternative ~100 usd + shipping:
https://enzymes.bio/product/beta-amylase-enzyme-powder-700000u-g-cas-9000-91-3/
125 g for ~16 usd + shipping.
https://onlineingredient.com/produc...-amylase-enzyme-powder?variant=41597769285677
Or a more expensive alternative ~100 usd + shipping:
https://enzymes.bio/product/beta-amylase-enzyme-powder-700000u-g-cas-9000-91-3/
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beren
Well-Known Member
Only said it because someone recommended it on Reddit for a too-high mash
OP
OP
Reddit...
The best choice for recovering from a too high mash is alpha amylase in the fermenter. Won't dry the beer out too much. On the other hand, if you want really dry, use amyloglucosidase (aka glucoamylase or just gluco) in the fermenter.
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You ain't kidding. I have about a gallon of what was 1.107 OG chocolate stout that is now 1.003 SG chocolate rocket juice (almost 14% ABV) courtesy of just a half teaspoon of Gluco I added a month ago - for SCIENCE!
Next, just for grins, I'm gonna see if CBC-1 can carbonate it in some PET liter bottles I have from my bottling days, but I can't imagine doing this to a full batch of...anything...for real.
Cheers!
